Relationship between cytokine single nucleotide polymorphisms and sarcoidosis among Japanese subjects.

Several susceptibility genes for sarcoidosis have been identified, but their relationship to the clinical state and prognosis remains to be elucidated. The aim of this study was to elucidate the relationship between sarcoidosis and five single nucleotide polymorphisms (SNPs) of three cytokines expected to play an important role in the inflammatory response. A case-control study was performed with 208 unrelated patients who met the diagnostic criteria for sarcoidosis used in Japan since 2006, and 328 control subjects. Five SNPs were analyzed: interleukin (IL)-10-819T/C (rs1800871), IL-10-592A/C(rs1800872), IL-6-634C/G (rs1800796), tumor necrosis factor-alpha (TNF-alpha)-857C/T (rs1799724), and TNF-alpha -1031T/C (rs1799964). No significant differences in SNPs were observed between the total sarcoidosis and control groups. However, the prevalence of rs1800871 and rs1800872 polymorphisms differed significantly in the sarcoidosis with eye involvement group compared with the control group [rs1800871 TT (vs. TC + CC): OR = 1.67, P = 0.034; rs1800872 AA (vs. AC + CC): OR = 1.66, P = 0.036]. Analyzing the cardiac involvement group, the prevalence of the rs1799724 polymorphism was significantly different from that of the control group [rs1799724 TT (vs. CC + CT): OR = 6.01. P = 0.006]. We concluded that the rs1799724 C/T polymorphism may affect susceptibility to cardiac sarcoidosis, while the rs1800871 T/C and rs1800872A/C polymorphisms may affect susceptibility to sarcoidosis with eye involvement.

Efficacy and safety of voriconazole in the treatment of chronic pulmonary aspergillosis: experience in Japan.

BACKGROUND: Though various clinical conditions of aspergillosis can occur, depending essentially on the host's immunological status, the focus of research in North American and European countries has mainly been on invasive pulmonary aspergillosis in immunocompromised patients. There are, however, also many problems to overcome in chronic forms of aspergillosis. One of those problems is that there are no codified treatment guidelines for chronic pulmonary aspergillosis (CPA). Especially in Japan, this issue is more serious, because there are more cases with CPA due to the many aged people with past history of tuberculosis. Several clinical cases and case series have reported the usefulness of the various antifungal agents that are available. The new triazole, voriconazole, in particular, seems to be effective in the treatment of CPA. The aim of the present study is to evaluate the efficacy and safety of voriconazole in the treatment of CPA in non-immunocompromised patients. PATIENTS AND METHODS: We conducted a prospective, open-label, non-comparative, multicenter study over a 2-year period. For inclusion in the study, patients with confirmed or probable CPA were recruited in 11 hospitals of the National Hospital Organization in Japan. Clinical, radiological, serological, and mycological data were collected at baseline and 12 weeks after treatment or at the end of treatment. RESULTS: Among 77 patients enrolled in the study, 71 patients (mean age 65.9 years, 56 males and 15 females) were eligible for the study. All of the eligible patients presented with underlying lung diseases, including sequelae of tuberculosis (n = 35), non-tuberculous mycobacterial lung disease (n = 8), chronic obstructive pulmonary disease (COPD) (n = 8), interstitial pneumonia (n = 7), cystic lung disease (n = 4), pneumothorax (n = 3), bronchial cancer (n = 1), and others (n = 5). Voriconazole was indicated in 48 cases (68 %) as the first-line treatment for CPA and 23 patients previously received other antifungal therapies. Based on a composite of clinical, radiologic, serological, and mycologic criteria, good response was seen in 43 patients (60.6 %), no response was observed in 19 patients (26.8 %), and 4 cases (5.6 %) got worse. Five patients (7.0 %) were unassessable for efficacy. The common adverse events were visual disturbances (17 patients, 23.9 %), abnormal liver function test results (12 patients, 16.9 %), adverse psychological effects (3 patients, 4.2 %), and others (10 patients, 14.0 %). Treatment with voriconazole had to be stopped in 2 cases (2.8 %) because of serious adverse events (abnormal liver function test results). There was no association between adverse effects and trough voriconazole levels in serum. CONCLUSIONS: In Japan, voriconazole provides effective therapy of CPA in non-immunocompromised patients with an acceptable level of toxicity.

Efficacy and outcomes of clarithromycin treatment for pulmonary MAC disease.

SETTING: Retrospective review of patients with pulmonary Mycobacterium avium complex (MAC) disease treated with clarithromycin. OBJECTIVES: To investigate whether the short-term response to treatment predicts long-term outcomes, and to analyse what explanatory variables are associated with the efficacy and outcome of treatment. DESIGN: Sputum conversion rates in short- and long-term outcomes were evaluated for 111 patients. Respectively 9 and 10 explanatory variables were analysed for their association with both response and outcomes. RESULTS: Eighty-four patients (75.7%) showed good short-term response and 94 (82.0%) showed good long-term outcomes. Women and patients with satisfactory nutrition status showed good short-term response. Patients with small lesions and those treated for >12 months after sputum conversion showed good long-term outcomes. Patients who showed good short-term response, in the group with large lesions, showed significantly good long-term outcomes (P = 0.0382). CONCLUSION: There were differences between prognostic factors reflecting short-term response and long-term outcomes. The short-term response predicts long-term outcomes in certain groups divided by prognostic factor. To establish standard treatment for pulmonary MAC disease, it is important to determine a standardised method of evaluation of treatment taking such factors into consideration.

bcl-2 and p53 protein expression in non-small cell lung cancers: correlation with survival time.

Apoptosis-related genes have received increasing attention in carcinogenesis, drug sensitivity, radiation sensitivity, and patient survival. bcl-2 and mutated p53 genes have been reported to inhibit apoptosis. To determine bcl-2 and p53 protein expression and their impacts on survival time in lung cancers, we studied 99 surgically resected, paraffin-embedded non-small cell lung cancer (NSCLC) specimens by immunohistochemical staining. The bcl-2 protein was expressed in 19.2% of NSCLCs. bcl-2-positive cases were found in 30. 4% of stages I and II carcinomas in 36.8% of squamous cell carcinomas. Patients with bcl-2 expression survived longer than those without. p53 protein was found in 44.4%; there was no significant difference in survival time between patients with and without p53 expression. Patients who were both bcl-2 positive and p53 negative survived significantly longer than those who were bcl-2 negative or p53 positive. These results suggest that bcl-2 protein expression can be histologically specific and stage dependent, and that the bcl-2 protein expression is potentially valuable for prognosis in NSCLC, particularly in the early stages, when bcl-2 protein expression is considered with mutant p53 protein expression.

Molecular epidemiology of tuberculosis in the north Hokkaido district of Japan.

SETTING: The incidence of tuberculosis in Japan has fallen rapidly in the past 50 years, to 27.9 cases per 100,000 population in 2001. OBJECTIVE: To assess the molecular epidemiology of tuberculosis in the north Hokkaido district, Japan. DESIGN: Restriction fragment length polymorphism analysis was consecutively performed on Mycobacterium tuberculosis strains from patients with pulmonary tuberculosis from April 1999 to March 2002. RESULTS: Of 229 patients, strains from 227 (99.1%) were available for analysis. The mean age of the patients analysed was 69.4 years. There was one immigrant patient. Two hundred and seven patients had six or more copies of IS6110. Of these, 16 (7.7%) in eight clusters had identical patterns, 120 (58.0%) belonged to five groups that had similar patterns (Dice coefficient >0.7) and 80 (38.6%) belonged to the 35 groups with the most common patterns (Dice coefficient >0.9). CONCLUSION: These results may reflect the epidemiological characteristics, age and migration of the residents and the incidence of tuberculosis of the area, and also those of Japan: many elderly patients were infected in the past, when the incidence of tuberculosis was very high.

Impaired interleukin-1beta converting enzyme (ICE) activity in patients with pulmonary tuberculosis.

We measured the activity of IL-1beta converting enzyme (ICE) of monocytes from pulmonary tuberculosis (PTB) patients. Peripheral blood mononuclear cells (PBMCs) and serum were collected from PTB patients (n = 16) and healthy subjects (n = 6). Serum IL-18 levels were significantly higher in PTB patients (515.9 +/- 326.5 pg/ml) than in controls (108.7 +/- 65.2 pg/ml) (P < 0.01), but gradually decreased with tuberculosis therapy. ICE activity was significantly lower in TB patients (65.3 +/- 34.4 vs. 142.2 +/- 75.6 U/mg) (P < 0.05). Serum IL-18 levels inversely correlated with ICE activity (P < 0.05). Altered ICE activity may explain why there is reduced IFN-gamma production from PBMCs; however, serum IL-18 is probably not regulated by ICE in monocytes in TB patients.

Disseminated Mycobacterium avium complex (DMAC) in an immunocompetent adult.

A female patient with multiple osteomyelitis and pulmonary Mycobacterium avium disease visited an orthopaedic clinic with back pain. Systemic bone scan showed multiple sites of increased radioactivity in the vertebral bodies, right scapula, femurs and ribs. M. avium was isolated from sputum and a sample aspirated from the right scapula. The route of infection was unknown as there was no history of trauma or surgery. HIV testing was negative. As there was no underlying immunological disease she was diagnosed as disseminated M. avium complex (DMAC) disease in an immunocompetent adult. Cytokine production on several stimuli from peripheral blood mononuclear cells was similar to that in pulmonary M. avium patients. Sequence analysis of IFN-gamma receptor revealed no nucleotide substitution. We detected serotypes 1, 2 and 4 from mycobacteria cultured from the right scapula, and conclude that this case could be the result of undetected immune deficiency and/or unrecognised virulence of the infecting isolate.

Epidermal growth factor receptor expression correlates with poor prognosis in non-small cell lung cancer patients with p53 overexpression.

To determine whether cancer patients with tumor suppressor gene abnormality survive for a shorter time when their growth was stimulated by growth factors, we examined 290 non-small cell lung cancer (NSCLC) specimens for p53 and epidermal growth factor receptor (EGFR) protein expressions using immunohistochemical staining. The distribution of cases by pathological stage of tumor was 155 cases of stage I, 30 cases of stage II, 96 cases of stage III and 9 cases of stage IV. Pathological types were 142 adenocarcinomas, 127 squamous cell carcinomas, 17 large cell carcinomas and 4 other types of malignancy. Immunohistochemical staining was performed on the formalin fixed, paraffin-embedded materials with monoclonal antibodies DO-7 and clone EGFR.133. positive staining for EGFR was seen in 124 (42.8%) cases. More EGFR positive cases were found in squamous cell carcinomas than in non-squamous cell carcinomas (p=0.0121). Staining for p53 protein was observed in 147 (50.7%) specimens. Multivariate proportional hazard model analyses revealed EGFR protein expression as a risk factor in the patients with NSCLC (p=0.0240). Patients negative for both EGFR and p53 survived for a longer period of time (p=0.0427).

[The effect of suramin on human lung cancer cell line in vitro].

Recently, several investigators reported the effects of suramin, a drug used in the treatment of trypanosomiasis, onchocerciasis, and an inhibition of the binding of some growth factors to cell surface receptors. In present study, therefore, we examined the effects of epidermal growth factor(EGF) and suramin on cell proliferation and cell cycle kinetics of an established cell line from human lung cancer(PC-13). EGF showed a stimulatory effect on cell proliferation of PC-13, suggesting EGF behaves as a growth factor of PC-13. On the other hand, suramin inhibited the stimulatory effect of EGF to PC-13 in a dose dependent manner. In addition, it was also observed that suramin inhibits the cell cycle progression from G0/G1 phase to S phase. These results indicate that suramin may withhold the cell proliferation and cell growth via suppression of the EGF cell stimulatory effects.

[Two cases of acute eosinophilic pneumonia--analyses of cytokines in BALF before and after treatment].

Acute eosinophilic pneumonia (AEP) has been well recognized as sudden onset respiratory failure characterized by an increased eosinophilic cell count in the bronchoalveolar lavage fluid (BALF). Although several lines of research have demonstrated that the Th2 cytokine network is likely to play a pivotal role in the development of AEP, that mechanism has not been clearly understood yet. Previous reports published in Japan demonstrated that the BALF CD4/CD8 ratio in patients with AEP ranged between 1.1 and 5.8. In this report, we describe 2 cases of AEP in which the CD4/CD8 ratios were 0.3 and 7.8 when present. We measured IL-4, IL-5, IL-10 and IFN gamma in the BALF before and after treatment in these patients. IL-5 decreased drastically after the treatment phase, while the other cytokines did not change much. We concluded that the IL-5 concentration in the BALF is probably related to the development of AEP. In addition, we also speculated that the CD4/CD8 ratio in BALF from patients with AEP may be affected whether a patient has an atopic background or not.

Suramin inhibits the growth of non-small-cell lung cancer cells that express the epidermal growth factor receptor.

The epidermal growth factor (EGF) is a potent growth factor that is believed to enhance the proliferation of cancer cells by a paracrine or autocrine mechanism. EGF transduces various signals and finally stimulates cell proliferation upon binding to cell surface receptors. Prevention of the association of this peptide with its receptors might lead to the development of new modalities for treatment of lung cancer. Several investigators have reported that suramin has antiproliferative activity against cancer cells that express EGF receptors (EGF-R), and that it acts by blocking the binding of the ligand to its receptor. In this study, we analyzed the antitumor effect of suramin using two lines of lung cancer cells (A549 and PC-13), which express EGF-R, and a variety of assays. Receptor-binding assays confirmed that A549 and PC-13 cells have cell surface receptors for EGF. Suramin inhibited the binding of EGF to these receptors. EGF and fetal bovine serum (FBS) stimulated the proliferation of cells, but suramin inhibited these effects in a dose-dependent fashion. Suramin at 200 microg/ml reduced the growth of A549 and PC-13 cells by 25 and 15%, respectively, in medium that contained 1% FBS. Paradoxically, the concentrations of suramin that inhibited cell proliferation were lower than those that were effective in inhibiting the binding of EGF to its receptor. Although expression of c-fos and c-myc mRNA increased when cells were stimulated by EGF or FBS, suramin at 200 microg/ml did not markedly alter such expression. Suramin partially blocked the EGF-induced progression of the cell cycle from the G0/G1 to the S phase. These results suggest that suramin partially blocks EGF signal transduction. Suramin probably inhibits cell proliferation by inhibiting intranuclear enzymes, as well as by partial blockage of EGF signal transduction.

[A case of asthma exacerbated by sulfite contained in betamethasone].

Adverse reactions to preservatives contained in food and medications have been recognized with increasing frequency. We have recently seen an asthmatic patient in whom wheezing and dyspnea increased after injection of betamethasone. The patient was a 26-year-old woman who had had bronchial asthma since 1980. She was treated at another hospital for moderate wheezing and betamethasone injections were given in May 1988. After this treatment her condition deteriorated acutely and she was transferred to our hospital. To confirm the possible relationship between betamethasone (sulfite) and asthmatic attack, provocation challenge tests were conducted. Intradermal skin test revealed an immediate positive reaction to sodium bisulfite at a concentration of 100 mg/L. Challenge test with increasing amounts of sodium bisulfite showed a 52% decrease in FEV1, 50 min after inhalation of 5 mg/ml solution. In addition, another inhalation challenge was conducted by use of an Astograph. Rrs immediately increased during inhalation of 10 mg/ml solution. These results suggested that sulfite contained in betamethasone preparation provoked exaggerated bronchospasm in this patient.

Extrinsic allergic alveolitis induced by the yeast Debaryomyces hansenii.

A 65-yr-old female developed cough, fever and dyspnoea following repeated exposure to a home ultrasonic humidifier. High-resolution computed tomography showed ground-glass opacity in both lung fields. Arterial blood gas analysis gave an oxygen tension of 8.38 kPa (63 Torr). Pulmonary function testing revealed restrictive ventilatory impairment with a reduction in the diffusing capacity. The diagnosis of extrinsic allergic alveolitis (EAA) was confirmed by radiographic findings, pathological evidence of alveolitis and reproductive development by a provocation test to the humidifier water. The yeast Debaryomyces Hansenii was the only microorganism cultured from the water of the humidifier. The double diffusion precipitating test and lymphocyte proliferative response was positive for an extract of D. Hansenii, providing evidence to incriminate this fungus. This is the first described case of EAA caused by D. Hansenii.

Depression of peripheral chemosensitivity by a dopaminergic mechanism in patients with obstructive sleep apnoea syndrome.

In the present study, respiratory drives to chemical stimuli and peripheral chemosensitivity were evaluated in patients with obstructive sleep apnoea (OSAS). The effects of oral administration of domperidone, a selective dopamine D2-receptor antagonist, were also examined, to study the respiratory effects of endogenous dopamine on peripheral chemoreceptors. Sixteen patients with OSAS and nine normal control subjects were studied. Respiratory responses to hypercapnia and hypoxia were measured using the rebreathing method and isocapnic progressive hypoxia method, respectively. The hypoxic withdrawal test, which measures the decrease in ventilation caused by two breaths of 100% O2 under mild hypercapnic hypoxic conditions (end-tidal oxygen and carbon dioxide tensions approximately 8.0 kPa and 5.3-6.7 kPa, respectively), was used to evaluate peripheral chemosensitivity. In the patients with OSAS, ventilatory responses to hypercapnia and hypoxia were significantly decreased compared with those of control subjects. Hypoxic withdrawal tests showed that peripheral chemosensitivity was significantly lower in patients with OSAS than in normal subjects. Hypercapnic ventilatory response and peripheral chemosensitivity were enhanced by administration of domperidone in the patients with OSAS, although no changes in either of these were observed in the control subjects. The hypoxic ventilatory response and peripheral chemosensitivity in the patients with OSAS were each significantly correlated with severity of hypoxia during sleep. These findings suggest that peripheral chemosensitivity in patients with obstructive sleep apnoea syndrome may be decreased as a result of abnormality in dopaminergic mechanisms and that the reduced chemosensitivity observed in patients with obstructive sleep apnoea syndrome may affect the severity of hypoxia during sleep.

[A case of asymptomatic paraganglioma diagnosed by abnormal shadow on chest X-ray].

A 65-year-old man was noted to have an abnormal shadow on chest X-ray at mass screening examination, and was seen at Douhoku Byoin National Sanatorium. Chest X-ray showed a homogeneous mass shadow and a lytic lesion of the right 5th rib with extra-pleural tumor sign at the right upper and middle lung fields. Retroperitoneal tumor of 5 cm in diameter was detected in the right adrenal area by ultrasonography. Because of increasing plasma levels of norepinephrine and dopamine, the diagnosis of malignant pheochromocytoma with rib metastasis was made. Histologically, chromogranin staining was positive in each resected specimen. The retroperitoneal tumor was separate from the right adrenal gland. We report a rare case of malignant paraganglioma which was asymptomatic in spite of increasing plasma levels of catecholamines.

[A case of obstructive sleep apnea syndrome with increased lung water content during sleep].

A 57-year-old man was admitted complaining of sleep disturbance. All night polysomnography showed a pattern of obstructive sleep apnea. We performed 201Tl scintigraphy to evaluate hemodynamic change and degree of stress on the right ventricle during sleep, and compared it with a 201Tl scintigram during wakefulness. We recognized 201Tl uptake by the lung in the 201Tl scintigram during sleep, but not during wakefulness. To determine the mechanism of 201Tl uptake by the lung during sleep, we measured lung water content during sleep by double indicator dilution method (Nihon Koden, NTV-1100). We recognized an increase of lung water content during sleep. We consider that the increase of lung water content during sleep is caused by sleep apnea, probably by hemodynamic change due to negative pleural pressure swings during sleep apnea.

[Association between sleep apnea syndrome and coronary artery disease].

Patients with sleep apnea syndrome often suffer from cardiovascular disease, but the incidence of coronary artery disease (CAD) with coronary spasm in these patients is not known. In the present study, 14 of 37 men with sleep apnea syndrome diagnosed by all-night polysomnography were suspected to also have CAD, based on the results of non-invasive clinical examinations. Coronary angiography confirmed the diagnosis of CAD either organic stenosis or coronary spasm in 8 of the 14 patients. Those 8 did not differ significantly from the 21 patients without CAD, with regard to coronary risk factors or to the severity of their sleep apnea (apnea index, 4% desaturation ratio, and nadir of SaO2). Eleven patients received intracoronary injections of acetylcholine, which induced coronary spasm in 4 (36.4%) and coronary contraction in 2 (18.2%). Coronary spasm was induced in 4 of the patients with CAD (50.0%). Although the pathophysiologic link between sleep apnea syndrome and CAD is still unclear, these results suggest that patients with this syndrome frequently suffer from CAD, particularly from coronary spasm.